This chapter is not intended to be a conventional review of gradient elution chromatography. Instead, the goal of this chapter is to provide practical strategies for large-scale separations using this method. Comprehensive reviews have provided its fundamentals and applications [1-4] for analytical purposes. In response to the increasing need for high purity bioproducts, advances in analytical liquid chromatography are being exploited for bioseparations [5]. Many of these bioproducts are proteins or other macro-molecules. However, most current theories and application strategies in gradient elution chromatography were developed for analytical purposes of small compounds, and they might not be appropriate for arge-scale separations of macro-molecules, which will be generally described in this section and in detail in the remaining sections of this chapter.